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Molecular and Cellular Biology, March 2000, p. 2129-2137, Vol. 20, No. 6
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Deadenylation and Poly(A)-Dependent Translational Repression Mediated by the Caenorhabditis elegans tra-2 3' Untranslated Region in Xenopus Embryos

Sunnie R. Thompson,1,dagger Elizabeth B. Goodwin,2 and Marvin Wickens1,*

Department of Biochemistry, University of Wisconsin---Madison, Madison, Wisconsin 53706-1569,1 and Department of Cell and Molecular Biology and Lurie Cancer Center, Northwestern University Medical School, Chicago, Illinois2

Received 22 September 1999/Returned for modification 9 November 1999/Accepted 20 December 1999

The 3' untranslated region (3'UTR) of many eukaryotic mRNAs is essential for their control during early development. Negative translational control elements in 3'UTRs regulate pattern formation, cell fate, and sex determination in a variety of organisms. tra-2 mRNA in Caenorhabditis elegans is required for female development but must be repressed to permit spermatogenesis in hermaphrodites. Translational repression of tra-2 mRNA in C. elegans is mediated by tandemly repeated elements in its 3'UTR; these elements are called TGEs (for tra-2 and GLI element). To examine the mechanism of TGE-mediated repression, we first demonstrate that TGE-mediated translational repression occurs in Xenopus embryos and that Xenopus egg extracts contain a TGE-specific binding factor. Translational repression by the TGEs requires that the mRNA possess a poly(A) tail. We show that in C. elegans, the poly(A) tail of wild-type tra-2 mRNA is shorter than that of a mutant mRNA lacking the TGEs. To determine whether TGEs regulate poly(A) length directly, synthetic tra-2 3'UTRs with and without the TGEs were injected into Xenopus embryos. We find that TGEs accelerate the rate of deadenylation and permit the last 15 adenosines to be removed from the RNA, resulting in the accumulation of fully deadenylated molecules. We conclude that TGE-mediated translational repression involves either interference with poly(A)'s function in translation and/or regulated deadenylation.

* Corresponding author. Mailing address: Department of Biochemistry, University of Wisconsin---Madison, 433 Babcock Dr., Madison, WI 53706-1569. Phone: (608) 262-8007. Fax: (608) 265-2603. E-mail: wickens{at}biochem.wisc.edu.

dagger Present address: Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305.

Molecular and Cellular Biology, March 2000, p. 2129-2137, Vol. 20, No. 6
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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